Optimal maize loci
First Claim
Patent Images
1. A maize plant, maize plant part, or maize plant cell comprising a recombinant nucleic acid molecule, said recombinant nucleic acid molecule comprising a nongenic maize genomic nucleic acid of at least 1 Kb determined to comprise the following characteristics:
- a. the level of methylation of said nongenic nucleic acid is 1% or less;
b. said nongenic nucleic acid shares less than 40% sequence identity with any other nucleic acid contained in the Zea mays genome;
c. said nongenic nucleic acid is located within a 40 Kb region of a known or predicted expressive maize coding nucleic acid; and
d. said nongenic nucleic acid exhibits a recombination frequency within the maize genome of greater than 0.00041 cM/Mb wherein said nongenic nucleic acid comprises a 1Kb nucleic acid having at least 95% sequence identity with a nucleic acid selected from the group consisting of SEQ ID NO;
1, SEQ ID NO;
100, SEQ ID NO;
203, SEQ ID NO;
295, SEQ ID NO;
384, SEQ ID NO;
687, SEQ ID NO;
781, SEQ ID NO;
843, SEQ ID NO;
967, SEQ ID NO;
1190, SEQ ID NO;
1252, SEQ ID NO;
1571, SEQ ID NO;
1663, SEQ ID NO;
1906, SEQ ID NO;
2027, SEQ ID NO;
2171, SEQ ID NO;
2256, SEQ ID NO;
2428, SEQ ID NO;
2632, and SEQ ID NO;
2649; and
a DNA of interest, wherein the DNA of interest comprises a non-native exogenous nucleic acid, and said DNA of interest is inserted into said nongenic nucleic acid.
2 Assignments
0 Petitions
Accused Products
Abstract
As disclosed herein, optimal native genomic loci have been identified in monocot plants, such as maize plants, that represent best sites for targeted insertion of exogenous sequences.
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Citations
10 Claims
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1. A maize plant, maize plant part, or maize plant cell comprising a recombinant nucleic acid molecule, said recombinant nucleic acid molecule comprising a nongenic maize genomic nucleic acid of at least 1 Kb determined to comprise the following characteristics:
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a. the level of methylation of said nongenic nucleic acid is 1% or less; b. said nongenic nucleic acid shares less than 40% sequence identity with any other nucleic acid contained in the Zea mays genome; c. said nongenic nucleic acid is located within a 40 Kb region of a known or predicted expressive maize coding nucleic acid; and d. said nongenic nucleic acid exhibits a recombination frequency within the maize genome of greater than 0.00041 cM/Mb wherein said nongenic nucleic acid comprises a 1Kb nucleic acid having at least 95% sequence identity with a nucleic acid selected from the group consisting of SEQ ID NO;
1, SEQ ID NO;
100, SEQ ID NO;
203, SEQ ID NO;
295, SEQ ID NO;
384, SEQ ID NO;
687, SEQ ID NO;
781, SEQ ID NO;
843, SEQ ID NO;
967, SEQ ID NO;
1190, SEQ ID NO;
1252, SEQ ID NO;
1571, SEQ ID NO;
1663, SEQ ID NO;
1906, SEQ ID NO;
2027, SEQ ID NO;
2171, SEQ ID NO;
2256, SEQ ID NO;
2428, SEQ ID NO;
2632, and SEQ ID NO;
2649; anda DNA of interest, wherein the DNA of interest comprises a non-native exogenous nucleic acid, and said DNA of interest is inserted into said nongenic nucleic acid. - View Dependent Claims (2, 3, 4, 5)
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6. A method of making a transgenic maize plant cell comprising a DNA of interest targeted to one nongenic maize genomic nucleic acid, the method comprising:
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a. selecting a nongenic maize genomic nucleic acid of at least 1 Kb, wherein said nongenic nucleic acid comprises the following characteristics; i). the level of methylation of said nongenic nucleic acid is 1% or less; ii). said nongenic nucleic acid shares less than 40% sequence identity with any other nucleic acid contained in the Zea mays genome; iii). said nongenic nucleic acid is located within a 40 Kb region of a known or predicted expressive maize coding nucleic acid; and iv). said nongenic nucleic acid exhibits a recombination frequency within the maize genome of greater than 0.00041 cM/Mb;
further wherein said nongenic nucleic acid comprises a 1 Kb nucleic acid having at least 95% sequence identity with a nucleic acid selected from the group consisting of SEQ ID NO;
1, SEQ ID NO;
100, SEQ ID NO;
203, SEQ ID NO;
295, SEQ ID NO;
384, SEQ ID NO;
687, SEQ ID NO;
781, SEQ ID NO;
843, SEQ ID NO;
967, SEQ ID NO;
1190, SEQ ID NO;
1252, SEQ ID NO;
1571, SEQ ID NO;
1663, SEQ ID NO;
1906, SEQ ID NO;
2027, SEQ ID NO;
2171, SEQ ID NO;
2256, SEQ ID NO;
2428, SEQ ID NO;
2632, and SEQ ID NO;
2649;b. introducing a site specific nuclease into a plant cell, wherein the site specific nuclease cleaves said nongenic nucleic acid; c. introducing the DNA of interest into the plant cell; d. targeting the insertion of the DNA of interest into said nongenic nucleic acid, wherein the cleavage of said nongenic nucleic acid facilitates integration of the DNA of interest into said nongenic nucleic acid; and e. selecting transgenic plant cells comprising the DNA of interest inserted into said nongenic nucleic acid. - View Dependent Claims (7, 8, 9, 10)
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Specification